title: A fusion protein between streptavidin and the endogenous TLR4 ligand EDA targets biotinylated antigens to dendritic cells and induces T cell responses in vivo.
abstract: The development of tools for efficient targeting of antigens to antigen presenting cells is of great importance for vaccine development. We have previously shown that fusion proteins containing antigens fused to the extra domain A from fibronectin (EDA), an endogenous TLR4 ligand, which targets antigens to TLR4-expressing dendritic cells (DC), are highly immunogenic. To facilitate the procedure of joining EDA to any antigen of choice, we have prepared the fusion protein EDAvidin by linking EDA to the N terminus of streptavidin, allowing its conjugation with biotinylated antigens. We found that EDAvidin, as streptavidin, forms tetramers and binds biotin or biotinylated proteins with a Kd ~ 2.6 × 10(-14) mol/L. EDAvidin favours the uptake of biotinylated green fluorescent protein by DC. Moreover, EDAvidin retains the proinflammatory properties of EDA, inducing NF- κβ by TLR4-expressing cells, as well as the production of TNF- α by the human monocyte cell line THP1 and IL-12 by DC. More importantly, immunization of mice with EDAvidin conjugated with the biotinylated nonstructural NS3 protein from hepatitis C virus induces a strong anti-NS3 T cell immune response. These results open a new way to use the EDA-based delivery tool to target any antigen of choice to DC for vaccination against infectious diseases and cancer.
authors.affiliation: Gene Therapy and Hepatology Area, Center for Applied Medical Research (CIMA), University of Navarra, 31008 Pamplona, Spain.
authors.forename: Laura
authors.initials: L
authors.lastname: Arribillaga
authors.forename: Maika
authors.initials: M
authors.lastname: Durantez
authors.forename: Teresa
authors.initials: T
authors.lastname: Lozano
authors.forename: Francesc
authors.initials: F
authors.lastname: Rudilla
authors.forename: Federico
authors.initials: F
authors.lastname: Rehberger
authors.forename: Noelia
authors.initials: N
authors.lastname: Casares
authors.forename: Lorea
authors.initials: L
authors.lastname: Villanueva
authors.forename: Marta
authors.initials: M
authors.lastname: Martinez
authors.forename: Marta
authors.initials: M
authors.lastname: Gorraiz
authors.forename: Francisco
authors.initials: F
authors.lastname: Borrás-Cuesta
authors.forename: Pablo
authors.initials: P
authors.lastname: Sarobe
authors.forename: Jesús
authors.initials: J
authors.lastname: Prieto
authors.forename: Juan José
authors.initials: JJ
authors.lastname: Lasarte
mesh_terms: D000818:Animals; D000938:Antigen-Presenting Cells / Q000276:immunology* / Q000378:metabolism; D002460:Cell Line; D003713:Dendritic Cells / Q000276:immunology / Q000378:metabolism; D005353:Fibronectins / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D016174:Hepacivirus / Q000276:immunology; D006801:Humans; D016233:Immunotherapy, Active; D008024:Ligands; D051379:Mice; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D019809:Streptavidin / Q000737:chemistry; D013601:T-Lymphocytes / Q000276:immunology / Q000473:pathology; D051197:Toll-Like Receptor 4 / Q000276:immunology* / Q000378:metabolism
chemical_list: D005353:Fibronectins; D008024:Ligands; D011993:Recombinant Fusion Proteins; C493487:Tlr4 protein, mouse; D051197:Toll-Like Receptor 4; D019809:Streptavidin